Process for obtaining ochropamine

ABSTRACT

The invention mainly pertains to a process for obtaining Ochropamine which comprises the steps of exhausting a vegetal drug by means of a solvent in an alkaline environment, separating the bases contained in the solvent by means of acid water, treating said acid water with a view to extracting therefrom the total alkaloids, and isolating the Ochropamine by chromatography, characterized in that said vegetal drug is the Cabucala Striolata Caudata.

United States Patent, [191 Hannart [451 Sept. 25, 1973 PROCESS FOROBTAINING OCHROPAMINE [75] inventor: Jean Alfred Alphonse Josephllannart, Brussels, Belgium [73] Assignee: Forschag Forschungsune ChemieAG, Basle, Switzerland 22 Filed: Sept. 24, 1971 21 Appl.No.: 183,693

52, [15.0. 260/29355 51 int. Cl. C07d 57/00 581 Field of Search..260/293.55

[56] References Cited OTHER PUBLICATIONS Douglas et al., Austral. J.Chem. l7, 246-55 (1964).

Zetler, Arzneimittel-Forsch. 14(12), 1277-86 (1964).

Primary Examiner-Henry R. Jiles Assistant ExaminerG. Thomas ToddAttorneyRichards & Geier ABSTRACT 1 Claim, N0 Drawings PROCESS FOROBTAINING OCHROPAMINE The present invention relates to a process forobtaining Ochropamine as well as the Ochropamine thus obtained,particularly as a therapeutic product.

Ochropamine, an indole alkaloid, has been extracted from the Ochrosia,particularly Ochrosia Poweri (see B. Douglas, J.L. Kirkpatrick,B.P.Moore and LA. Weisbach, Austral J.Chem.l7, 246, 1964).

The Applicant has found that Ochropamine has antiviral properties,whereas its toxicity is very weak.

On the other hand, the Applicant has also found, that Ochropamine couldbe extracted from Cabucala, an Apocynacea of the tropical regions, andmore particularly form the Cabucala Striolata Caudata. The .part of thisplant having the largest content are its leaves, from dium sulfate,filtered and evaporated until dry. Thus which, according to the samples,from 8 to 10.5 g. total alkaloids per kg dry leaves may be extracted,out of which 2 to 3.5 g of pure Ochropamine can be isolated.

The process according to the present invention comprises the steps ofexhausting a vegetal drug by means of a solvent in an alkalineenvironment, separating the bases contained in the solvent by means ofacid water, treating said acid water with a view to extracting therefromthe total alkaloids, and isolating the Ochropamine by chromatography,said process being characterized in that said vegetal drug is theCabucala Striolata Caudata. Preferably the leaves of the latter areused.

The Ochropamine thus obtained clearly exhibits antiviral properties ontobacco mosaic and on the influenza virus. On the otherhand thefollowing pharmacodynamic properties may be noted: Toxicity on miceintravenous administration DL 50 39 mg/kg.

With cats, acceleration of the respiratory rhythm and increasedamplitude.

Decrease of fatigability with the rat.

Increased cursiosity mice.

Slight increase of the arterial tension for 1/5th of the DL 50.

Small effect on the rhythm and amplitude of the heart.

An example to illustrate how the invention is to be carried into effectwill be described in detail hereinafter.

Leaves of Cabucala Striolata are dried at the places of harvest, inthick layers, frequently stirred, in the shadow. It is also possible todry the leaves in drying kilns ventilated with hot air. The drug isconsidered to be dry, when its water content has dropped to 12 percent.

The dry leaves are ground and 4 kg is weighed out. The powder is thenalkalized by kneading it with 4 l of a watery solution of 10 percentsodium carbonate hydrate, then left to rest for 2 hours. The vegetalmass is then introduced into a Soxhlet type of extracting apparatus andexhausted during three hours by means of ethyl ether. At the end of thistreatment 27.5 1 of ether is obtained. This solvent is now concentratedso as to reduce its volume to 6 1. Then after filtration it is exhaustedby 6 washings each of 500 ml of phosphoric water of 1 percent. In thisway about 3 1 water containing all the bases as phosphates is obtained.This water is stripped in vaccum of residual quantities of solvent, thenstirred with 50 g of bone-black and filtered. After gradual alkalizationby means of ammonia, the bases are extracted by stirring the aqueousphase with 250 ml of chloroform. 6 washings are effected with thissolvent, then are joined, washed with water, dried on soone obtains 40 gtotal alkaloids, Le. a yield of 10 g. per kg dry leaves.

The total alkaloids are dissolved in benzene and passed on a column of1,500 g alumina. The following series of solvents is used for elution:

pure benzene benzene/ether 50-50 pure ether chloroform chloroform with1% of methyl alcohol chloroform with 1.5% of methyl alcohol TheOchropamine is obtained in the chloroform fractions with low methylalcohol content. The chromatography can be continued in order to obtainthe other alkaloids, by increasing the methyl alcohol content of thechloroform.

The fractions containing the Ochropamine are joined and evaporated untilthey are dry. A residue of 17.5 g. is thus obtained. This is taken upagain with ethyl alcohol en by concentration of the solvent yields afirst cast of 8.15 g. and a second cast of 2.1 g, i.e. 10.25 gOchropamine in total.

The Ochropamine thus obtained has the following confirmatorycharacteristics:

1. Melting point: 134 C.

2. Rotating power (a),,= 153(acetone) 3. Molecular formula C,,H,,0,N,

4. Molecular weight 366.44

5. Centesimal compositiomC 72.10

OCH3 8.46

6. U.V.Spectrum 243(4.28) 315(4.25)

7. l.R.Spectrum 3,400; 2,940; 2,770; 1,730; 1,715; 1,650; 1,615; 750

8. R.M.N.Spectrum: one quartet of 3 protons centered on 1.7

one singlet of 6 protons at 2.55;

one singlet of 3 protons at 4.03;

one quartet of 1 proton centered on 5.4

fou'r aromatic protons between 7 and 8 ppm.

9. Mass spectrography: M -peak at 366 and principal peaks at 351,334-335 323,307, 252, 222, 188, 180, 172,143,134,122,120,115.

In consideration of these characteristics it may be concluded that thesubstance obtained really is Ochropamine, having the structural formula:

CHaOOC H l/CH: N N

O W CH3 cess to isolate the Ochropamine, elution in said chromatographicprocess being successively carried out with pure benzene, a mixture of50 50 benzene and ether, pure ether, chloroform, chloroform with lpercent methyl alcohol added and chloroform with 1.5 percent methylalcohol added, Ochropamine being obtained in the chloroform fractionswith a low methyl a1- cohol content.

UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,761,13} Dated September 25', 1973 Invent0r(s) Jean Alfred Alphonse JosephHannart It is certified that error appears in the above-identifiedpatent and that said Letters Patent are hereby corrected as shownbelow:-

On the cover sheet item should. read as follows:

-- FORSGHAG, FORS"I NGSUND CHEMIE AG, Basle, Switzerland Signed andsealed this 30th day of April 19714..

(SEAL) Attest:

EDWARD I-T.FLETCHER,JR G. MARSHALL DANN Attesting Officer Commissionerof Patents

